Question

Design for introducing batch effect in DESeq2

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ag1805x ▴ 80

@ag1805x-15215

Last seen 12 weeks ago

University of Allahabad

I have RNAseq data taken from two different ENA project IDs (PrjA & PrjB). Both PrjA & PrjB contains test and control samples. PrjA was PE 50bp sequencing while PrjB was SE 100bp. Is it possible we can combine the data to identify DEG between test and control? Would batch effect removal by Combat be a better option or use the design = project+condition in DESeq2?

deseq2 combat batch effect rna-seq edger • 1.0k views

ADD COMMENT • link updated 4.7 years ago by swbarnes2 ★ 1.3k • written 4.7 years ago by ag1805x ▴ 80

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If you DO know the batches, then what swbarnes2 recommended is the best option. If you don't know then you could use sva.

In your case, however, I don't think that it's wise to just combine the data because it seems that you use different platforms for the sequencing. Could you show us the MDS plot?

ADD REPLY • link 4.7 years ago mikhael.manurung ▴ 270

score 1 · Answer 1 · 2019-08-30

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swbarnes2 ★ 1.3k

@swbarnes2-14086

Last seen 26 minutes ago

San Diego

Do what the DESeq software recommends. Give DESeq raw counts, not some kind of batch corrected, and include batch as a factor in the design.

ADD COMMENT • link 4.7 years ago swbarnes2 ★ 1.3k

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You can do that even when the data came from two different projects? Seems like the batch issue in this case is not simply multiple runs of the same instrument but other things are different as well.

ADD REPLY • link 4.7 years ago mikhael.manurung ▴ 270